【摘要】 目的:研究siRNA对人骨肉瘤U20S细胞MDM2基因表达及肿瘤细胞增殖的抑制作用。方法:构建可表达针对MDM2的siRNA质粒(PGCsilencerTM-MDM2-siRNA)。转染siRNA MDM2(简称siMDM2),阴性对照质粒到U20S,并设只加转染试剂作空白对照组,用逆转录聚合酶链反应(RT-PCR)和Western blotting法检测siMDM2对MDM2基因和蛋白表达的抑制作用,并用MTT法检测siMDM2对细胞增殖的抑制作用。结果:RT-PCR结果显示,转染siMDM2-1和-2组MDM2的mRNA表达量分别下调到空白对照组的32.61%和39.06%;Western blotting结果显示,转染siMDM2-1和-2组蛋白表达量下调到空白对照组的35.76%和42.20%;转染阴性对照质粒的MDM2基因和蛋白与转染试剂组比较差异均无显著性(P>0.05)。MTT结果显示,转染siMDM2后细胞生长受到明显抑制,与转染试剂组比较差异具有显著性(P<0.05),阴性对照组抑制率与转染试剂组比较差异无显著性(P>0.05)。结论:siRNA可以有效地抑制U20S细胞中MDM2的表达,并抑制细胞增殖。更多还原

【Abstract】 Objective To study the inhibitory effects of siRNA on MDM2 expression in osteosarcoma cell line U20S and cell proliferation.Methods siRNA targeting MDM2(PGCsilencerTM-MDM2-siRNA) was constructed and transfercted into U20S cells.Meanwhile,control siRNA was transfected and the transfection agent was used as the comparison.MDM2 gene and protein expressions were measured using RT-PCR and Western blotting.Cell proliferation was measured with MTT assay.Results RT-PCR and Western blotting results showed that MDM2 mRNA expression levels were reduced to 32.61% and 39.06% after being transfected with siMDM2-1 and siMDM2-2; MDM2 protein expression levels decreased to 35.76% and 42.20%,respectively.The difference between control siRNA and liposome group was not obvious(P>0.05).MTT result showed that the cell growth was inhibited by being transfected with siMDM2,the difference was obvious compared with liposome group(P<0.05).The difference was not significant between control siRNA and liposome group (P>0.05).Conclusion siRNA can effectively down-regulate MDM2 expression in U20S cell line and inhibit the cell proliferation.更多还原